The present invention generally relates to apparatus and methods for performing real time assays and specifically for performing real time PCR in array tape.
An endpoint assay performs a single measurement after a reaction is complete, while a real time assay performs multiple measurements during the reaction. Measuring a reaction over time is known as ‘chemical kinetics’ or ‘reaction kinetics’. This kinetic data provides additional information about the reaction process.
The benefits of real time assays over endpoint assays are many. Chemical reactions often follow a natural exponential rate. By measuring the reaction kinetics, the exponential rate may be observed and quantified. The amount of initial reagent may be quantified based upon the kinetics. The kinetics of one reaction may be compared with a similar reaction or a standardized reaction.
Polymerase Chain Reaction (PCR) causes DNA amplification. The measure of the reaction is based upon measuring the amount of DNA in the sample. This is typically done using fluorescent probes. Some PCR chemistries incubate using a single reaction temperature, while others use multiple temperatures. When using multiple temperature incubation, ideally each strand of DNA will be limited to doubling with each thermal cycle. In the case of single temperature reactions, the DNA amplification will proceed at some rate, and measurements would be taken periodically.
Although real time assays have been performed utilizing plates and tubes, such real time assays are conventionally not performed on large scale experiments due to various factors including cost, speed, and the like.
Thus, a need exists for apparatus and methods for performing real time assays which overcome the deficiencies and shortcomings of conventional apparatus and methods of doing so.